Anti-FACL4 抗体 [EPR17587]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-FACL4 antibody [EPR17587] (AB205197)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma) cells labeling FACL4 with ab205197 at 1/50 dilution followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HepG2 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows :
-ve control 1 : ab205197 at 1/50 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FACL4 antibody [EPR17587] (AB205197)

Immunohistochemical analysis of paraffin-embedded Human liver carcinoma tissue labeling FACL4 with ab205197 at 1/250 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on Human hepatocellular carcinoma tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-FACL4 antibody [EPR17587] (AB205197)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling FACL4 with ab205197 at 1/50 dilution followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows :
-ve control 1 : ab205197 at 1/50 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

• WB

Lab

Western blot - Anti-FACL4 antibody [EPR17587] (AB205197)

Western blot : Anti-FACL4 antibody [EPR17587] ab205197 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 79 kDa in Wild-type A431 cell lysates with no signal observed at this size in ACSL4 knockout A431 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-FACL4 antibody [EPR17587] (ab205197) at 1/1000 dilution

Lane 1:

Wild-type A431 cell lysates at 20 µg

Lane 2:

Western blot - Human ACSL4 (FACL4) knockout A-431 cell line (<a href='/products/cell-lines/human-acsl4-facl4-knockout-a-431-cell-line-ab269613'>ab269613</a>) at 20 µg

Lane 3:

HEK-293 cell lysates at 20 µg

Lane 4:

K562 cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 79 kDa

Observed band size: 79 kDa

false

• WB

Supplier Data

Western blot - Anti-FACL4 antibody [EPR17587] (AB205197)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-FACL4 antibody [EPR17587] (ab205197) at 1/1000 dilution

All lanes:

293 (Human epithelial cells from embryonic kidney) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 79 kDa

Observed band size: 79 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-FACL4 antibody [EPR17587] (AB205197)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-FACL4 antibody [EPR17587] (ab205197) at 1/1000 dilution

All lanes:

Human fetal kidney tissue lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 79 kDa

Observed band size: 79 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-FACL4 antibody [EPR17587] (AB205197)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-FACL4 antibody [EPR17587] (ab205197) at 1/5000 dilution

Lane 1:

HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

HepG2 (Human liver hepatocellular carcinoma) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 79 kDa

Observed band size: 79 kDa

false

Exposure time: 3min